LOCUS       sequence-384099-        7154 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector to drive the Flp-dependent expression of mEGFP (L221K) 
            -Gphn (isoform P1) under the control of human Synapsin promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7154)
  AUTHORS   Liebsch F, Eggersmann FR, Merkler Y, Kloppenburg P, Schwarz G
  TITLE     Automated Image Analysis Reveals Different Localization of Synaptic 
            Gephyrin C4 Splice Variants.
  JOURNAL   eNeuro. 2023 Jan 4;10(1):ENEURO.0102-22.2022. doi: 
            10.1523/ENEURO.0102-22.2022. Print 2023 Jan.
  PUBMED    36543537
REFERENCE   2  (bases 1 to 7154)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1523/ENEURO.0102-22.2022"; journalName: "eNeuro"; date: 
            "2023-01-4- 4"; volume: "10"; issue: "1"; pages: 
            "ENEURO.0102-22.2022"
COMMENT     Sequence Label: pAAV-hSyn-fDIO-mEGFP-Gphn_P1
FEATURES             Location/Qualifiers
     source          1..7154
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     40..61
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(215..234)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     334..356
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(394..412)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        480..584
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(585..653,654..1445)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      1616..2204
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2105..2124
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   2266..2395
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        2440..2887
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    2925..2958
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     CDS             complement(join(5286..5996,5997..5999,6000..6002))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLK
                     EFVTAAGITLGMDELYK"
     primer_bind     5677..5696
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5936..5957
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      5999..6008
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(6015..6048)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     polyA_signal    6166..6642
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(6365..6384)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   6682..6822
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   6682..6811
                     /label=AAV2 ITR
     rep_origin      join(6897..7154,1..198)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6984..7003)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
       61 gaacaacact caactctatc tcgggctatt cttttgattt ataagggatt ttgccgattt
      121 cggtctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
      181 tattaacgtt tacaatttta tggtgcactc tcagtacaat ctgctctgat gccgcatagt
      241 taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
      301 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
      361 caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg
      421 ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc
      481 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
      541 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
      601 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
      661 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
      721 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
      781 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
      841 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
      901 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
      961 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     1021 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     1081 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     1141 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     1201 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     1261 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     1321 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     1381 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     1441 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     1501 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     1561 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     1621 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     1681 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     1741 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     1801 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     1861 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     1921 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     1981 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     2041 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     2101 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     2161 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     2221 cctttttacg gttcctggcc ttttgctggc cttttgctca catgtcctgc aggcagctgc
     2281 gcgctcgctc gctcactgag gccgcccggg cgtcgggcga cctttggtcg cccggcctca
     2341 gtgagcgagc gagcgcgcag agagggagtg gccaactcca tcactagggg ttcctgcggc
     2401 cgcacgcgtg tgtctagact gcagagggcc ctgcgtatga gtgcaagtgg gttttaggac
     2461 caggatgagg cggggtgggg gtgcctacct gacgaccgac cccgacccac tggacaagca
     2521 cccaaccccc attccccaaa ttgcgcatcc cctatcagag agggggaggg gaaacaggat
     2581 gcggcgaggc gcgtgcgcac tgccagcttc agcaccgcgg acagtgcctt cgcccccgcc
     2641 tggcggcgcg cgccaccgcc gcctcagcac tgaaggcgcg ctgacgtcac tcgccggtcc
     2701 cccgcaaact ccccttcccg gccaccttgg tcgcgtccgc gccgccgccg gcccagccgg
     2761 accgcaccac gcgaggcgcg agataggggg gcacgggcgc gaccatctgc gctgcggcgc
     2821 cggcgactca gcgctgcctc agtctgcggt gggcagcgga ggagtcgtgt cgtgcctgag
     2881 agcgcagtcg agaaggtacc ggatcctcta gagtcgactc cggagaagtt cctattctct
     2941 agaaagtata ggaacttcgc agaatggtag ctggattgta gctgctatta gcaatatgaa
     3001 acctcttaga agttcctatt cttcaaaagg tataggaact tcggcgcgcc tcatagccgt
     3061 ccgatgacca tgacgtccac cacctcgcct ttgtgaagtt ccacatactg ctctgtcttt
     3121 ggaggtagca tcagcaatcc attggcactg cgcatgctca tcagacggct gctcatctga
     3181 ttacctgtac tctgggccca gggcagtggt tcttggtgat gccaagtcag tatacaccgg
     3241 tggtattctg ggcgagggtc cagctttacg tcacatgata acctagcttt gatgatggtt
     3301 ggccgaggat ccaagatacc ctgcattttt ctcagtgcgg gtacaacaaa gaggttgcag
     3361 gtgaccacag ctgatacagg attccctggt agtgcaaaaa taatttttct tacaccatca
     3421 atatccaaag ttgcaaatgt tgttggcaag cctggcttca taaaaaccct gccaaaatgg
     3481 atctgagcat gaagatcgat gtccagcacc tgcttgagat agtccttttc ccccatggat
     3541 acacctcctg acgtgatgat gacatcagca cggctgatac cctcattcaa ggcattgagt
     3601 aagtcatctg ggttgtctcc tacaattccc aagttgattg ttgggtaacc atgttcctga
     3661 attgttgcta gaagtgttga tcgattgctg tcccgaatct ttcctggtaa gaggtcatct
     3721 tcaggattta gtagctcatt ccctgttgac ataacggcaa ccactggaaa cttattaact
     3781 tcaacttctg tgacacctac agttgcaagg agaccaattt ctgaagggcc catatgggtt
     3841 ccttttgcca aaacacactc ccctctttta atgtcgtggc cgatgggtct gatgtcttgg
     3901 cctggccgag cttgcaccag aattcgcact tcaagttctt cagtaccatc atctgattcc
     3961 ctgatgagtt cagtatcttc aacttgcact actgcatcag caccgcaggg gattggagca
     4021 cctgttgtaa cccgcatcac ttgtccgggc attactgtct gagttggctg ttcaccagct
     4081 tgggattccc caatgatgaa acgatctcct ggaccatcag cagctcgaac agcatagcca
     4141 tcttttactg atgctgggaa tgggggtagg ttgtcttttg catacacatc ttgagcaagg
     4201 actcgcccca ttccatctcg gtaattgatg atttctgtac caagcaccgg agtcatctcc
     4261 agaactgtaa tgaaggcttt gtccatagac gtcagaggaa aaggagacat gcgatgtctt
     4321 ctagccacct tggtgatatc tacagcactg tgactggctc ttagaatgtt ctccttgctg
     4381 ctgcacctgg actggacttt tggtgttgga caagaagctg tagagaggcg agatgtagcc
     4441 tgggcacggg gcgactctga aggagtagtg ctaagggagg ctgtgtctcg tgggagcacc
     4501 tgaacgccac gagaaatgat ggagtctgga atctttgcag caagagctgc tgcagtaata
     4561 tgtgatgagg aactatcttc tgtcgaagct acaccactgt ctttcttttc ttcttcctcc
     4621 tcttcacact gaactccttt gtcttctgtc tgcttatgtg ggctagttgt aggaggtgga
     4681 gaaagaggag gtggtgggga aggtaaatct tcaagttcat catgcacctc ctttactttt
     4741 acaatggcat cacgtaaaag gtcaatggca tgaggtagag ctggcagtat gaactgaaag
     4801 cattcctgag atcctttctt gctaccaggc aggttaatta tcagagtttt ccctcttatt
     4861 ccacacactg gtctagagag catgcccaga ggtgtaacat taagtgatcc catcagcatt
     4921 gccagggcca tccctggcgc ttcccgttct attacttctt ttgtggcctc tggagtgaca
     4981 tctcgtggtg caaaccctgt tccgccagtt gttaatatca aattaagttc cttctcatca
     5041 caccagtcta tgagtgtttc cttgatttct tctatttcat ctggtactat cttgtatgct
     5101 gatatagtcc cacccaacaa agaaggatcc tggacgagat cttttagatt tatcccactg
     5161 cggtcttctg caagattcct gaagcagcta tcactcactg tgaggactcc gacacggatt
     5221 tgatggtcgt ggtttgtgag gatcattccc tcggtcgcca tgctcgagat ctgagtccgg
     5281 ccggacttgt acagctcgtc catgccgaga gtgatcccgg cggcggtcac gaactccttc
     5341 aggaccatgt gatcgcgctt ctcgttgggg tctttgctca gggcggactg ggtgctcagg
     5401 tagtggttgt cgggcagcag cacggggccg tcgccgatgg gggtgttctg ctggtagtgg
     5461 tcggcgagct gcacgctgcc gtcctcgatg ttgtggcgga tcttgaagtt caccttgatg
     5521 ccgttcttct gcttgtcggc catgatatag acgttgtggc tgttgtagtt gtactccagc
     5581 ttgtgcccca ggatgttgcc gtcctccttg aagtcgatgc ccttcagctc gatgcggttc
     5641 accagggtgt cgccctcgaa cttcacctcg gcgcgggtct tgtagttgcc gtcgtccttg
     5701 aagaagatgg tgcgctcctg gacgtagcct tcgggcatgg cggacttgaa gaagtcgtgc
     5761 tgcttcatgt ggtcggggta gcggctgaag cactgcacgc cgtaggtcag ggtggtcacg
     5821 agggtgggcc agggcacggg cagcttgccg gtggtgcaga tgaacttcag ggtcagcttg
     5881 ccgtaggtgg catcgccctc gccctcgccg gacacgctga acttgtggcc gtttacgtcg
     5941 ccgtccagct cgaccaggat gggcaccacc ccggtgaaca gctcctcgcc cttgctcacc
     6001 atggtggcgc tagcgaagtt cctatacttt ctagagaata ggaacttctt gccttaaccc
     6061 agaaattatc actgttattc tttagaatgg tgcaaagaga agttcctata ccttttgaag
     6121 aataggaact tcgaattcga tatcaagctt atcgatgaga tctacgggtg gcatccctgt
     6181 gacccctccc cagtgcctct cctggccctg gaagttgcca ctccagtgcc caccagcctt
     6241 gtcctaataa aattaagttg catcattttg tctgactagg tgtccttcta taatattatg
     6301 gggtggaggg gggtggtatg gagcaagggg caagttggga agacaacctg tagggcctgc
     6361 ggggtctatt gggaaccaag ctggagtgca gtggcacaat cttggctcac tgcaatctcc
     6421 gcctcctggg ttcaagcgat tctcctgcct cagcctcccg agttgttggg attccaggca
     6481 tgcatgacca ggctcagcta atttttgttt ttttggtaga gacggggttt caccatattg
     6541 gccaggctgg tctccaactc ctaatctcag gtgatctacc caccttggcc tcccaaattg
     6601 ctgggattac aggcgtgaac cactgctccc ttccctgtcc ttctgatttt gtaggtaacc
     6661 acgtgcggac cgagcggccg caggaacccc tagtgatgga gttggccact ccctctctgc
     6721 gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc
     6781 gggcggcctc agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt
     6841 ttctccttac gcatctgtgc ggtatttcac accgcatacg tcaaagcaac catagtacgc
     6901 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     6961 acttgccagc gccttagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     7021 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     7081 tttacggcac ctcgacccca aaaaacttga tttgggtgat ggttcacgta gtgggccatc
     7141 gccctgatag acgg
//
